Validated using a knockout cell line

Anti-Nestin antibody [2C1.3A11] (ab18102)

Reviews (1)Q&A (1)References (9)

Key features and details

  • Mouse monoclonal [2C1.3A11] to Nestin
  • Suitable for: ICC/IF, WB, Flow Cyt, IHC-P, IP
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Nestin antibody [2C1.3A11]
    See all Nestin primary antibodies

  • Description

    Mouse monoclonal [2C1.3A11] to Nestin

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human
    Does not react with: Rat

  • Immunogen

    A 150 amino acid fragment from the cloned human Nestin.

  • General notes


    This highly specific antibody to human nestin can aid in characterizing progenitor cells differentiating into distinct lineages, in enhancing the therapeutic potential of human neural stem/progenitor cells in the treatment of CNS diseases or injury, and in identifying neuroepithelial tumor cells.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab18102 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/5000. Predicted molecular weight: 177 kDa.
Flow Cyt 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • Function

    Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells.

  • Tissue specificity

    CNS stem cells.

  • Sequence similarities

    Belongs to the intermediate filament family.

  • Developmental stage

    Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.

  • Post-translational
    modifications

    Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
  • Information by UniProt

  • Database links

    • Alternative names

      • ESTM 46 antibody
      • FLJ 21841 antibody
      • FLJ21841 antibody
      • Intermediate filament protein antibody
      • Nbla00170 antibody
      • nes antibody
      • NEST_HUMAN antibody
      • Nestin antibody
      see all

    Images

    • Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      All lanes : Anti-Nestin antibody [2C1.3A11] (ab18102) at 1/5000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : NES knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 177 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab18102 observed at 300 kDa. Red - loading control, ab176560, observed at 50 kDa.

      ab18102 was shown to recognize NES in wild-type HAP1 cells as signal was lost at the expected MW in NES knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NES knockout samples were subjected to SDS-PAGE. Ab18102 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunofluorescence staining of U251 cells (glioblastoma cell line) stained using ab18102(green) and bisbenzimide (blue)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of migrating tumor cells labelling Nestin with ab18102 at 2µg/ml. Cytoplasm and membrane positive staining shown.

    • Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)
      All lanes : Anti-Nestin antibody [2C1.3A11] (ab18102) at 1/1000 dilution

      Lane 1 : Total rat brain protein at 5 µg
      Lane 2 : Human CNS progenitor cell protein at 1 µg
      Lane 3 : Human CNS progenitor cell protein at 5 µg

      Predicted band size: 177 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of xenograft tumor cells labelling Nestin with ab18102.

    • Flow Cytometry - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Flow Cytometry - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Overlay histogram showing SH-SY5Y cells stained with ab18102 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18102, 1/200 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)
      Immunohistochemistry of Anti-Nestin on formalin fixed paraffin embedded prostate nerve tissue using ab18102

    References (9)

    Publishing research using ab18102? Please let us know so that we can cite the reference in this datasheet.

    ab18102 has been referenced in 9 publications.

    • Ashraf S  et al. The polyadenylation inhibitor cordycepin reduces pain, inflammation and joint pathology in rodent models of osteoarthritis. Sci Rep 9:4696 (2019). PubMed: 30886197
    • Yamagishi A  et al. The Structural Function of Nestin in Cell Body Softening is Correlated with Cancer Cell Metastasis. Int J Biol Sci 15:1546-1556 (2019). PubMed: 31337983
    • Ferrer CM  et al. An inactivating mutation in the histone deacetylase SIRT6 causes human perinatal lethality. Genes Dev 32:373-388 (2018). PubMed: 29555651
    • Lin Y  et al. Isoliquiritigenin inhibits the proliferation and induces the differentiation of human glioma stem cells. Oncol Rep 39:687-694 (2018). PubMed: 29251326
    • Zhao J  et al. APOE e4/e4 diminishes neurotrophic function of human iPSC-derived astrocytes. Hum Mol Genet 26:2690-2700 (2017). PubMed: 28444230
    • Wang Y  et al. Sevoflurane anesthesia in pregnant rats negatively affects nerve function in offspring potentially via inhibition of the Wnt/ß-catenin pathway. Mol Med Rep 15:2753-2759 (2017). PubMed: 28447764
    • Jaraíz-Rodríguez M  et al. A Short Region of Connexin43 Reduces Human Glioma Stem Cell Migration, Invasion, and Survival through Src, PTEN, and FAK. Stem Cell Reports 9:451-463 (2017). PubMed: 28712848
    • Sherry MM  et al. STAT3 is required for proliferation and maintenance of multipotency in glioblastoma stem cells. Stem Cells 27:2383-92 (2009). WB ; Human . PubMed: 19658181
    • Messam CA  et al. Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody. Exp Neurol 161:585-96 (2000). PubMed: 10686078

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Flow Cytometry abreview for Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker

     Excellent
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Neural Progenitor cells (from human ES cells))
    Specification
    Neural Progenitor cells (from human ES cells)
    Preparation
    Cell harvesting/tissue preparation method: Cell Dissociation Buffer
    Sample buffer: FACS buffer (0.2%BSA, 0.1%sodium azide in PBS)
    Fixation
    Paraformaldehyde
    Permeabilization
    Yes - 70% Methanol (ice cold)
    Gating Strategy
    FSC vs SSC on negative control sample
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 03 2012

    Question

    Yes, that would be for the nestin antibodies only. (ab6320 & ab18102) If you can find any specific FACS protocol for one of these, then it may be decisive in my choice. Up to know, I do not have any information for choosing between them. Thus I am waiting for your answer. Anyway, if you cannot find any protocols,then I will choose one of them by chance. Thanks in advance, I hope this helps

    Read More
    Answer

    I am please to be able to share the following protocol for ab18102for Flow cytometry: Fix cells with 4% Paraformaldehyde for 30 min at room temperature (RT) Wash twice with 1% PBS at 1500rpm/5min/RT Permeabilize cells with 0.02% Triton X-100 in 1%PBS + 1% BSA for 15 min/RT Then add rabbit nestin ab (1:400 dilution) for 1hr/RT Wash twice Add specific secondary ab for 1hr/RT Wash twice Flow reading I hope this information will be helpful. Please do not hesitate to contact me if I can be of further assistance,

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